The purpose of this study is to systematically study and analyze the dose calibration and measurement of the alanine/ESR dosimetry system in FLASH radiotherapy.The dose calibration curve of the alanine dosimeter ranged from 0 to 50 Gy, and the best linear fit to the data shows a correlation coefficient close to 1.In the stability experiment, the preset irradiation values for the alanine dosimeter and radiochromic film were 14.34 Gy and 13.38 Gy, resp., and the results of a stability experiment results showed that the alanine dosimeter exhibited superior stability, compared with radiochromic film, with a deviation of less than 1% in repeated measurements.The results of X-ray FLASH irradiation experiments on mice showed that the maximum standard errors of the alanine dosimeter between the measured results and the preset dose values were 4.57% (chest experiment) and 3.22% (abdomen experiment), whereas those for the radiochromic film were 6.03% (chest experiment) and 6.50% (abdomen experiment).Addnl., X-ray and electron-beam FLASH radiotherapy experiments on mice verified that the alanine dosimeter showed no dose-rate dependence and provided more accurate measurement results compared to the radiochromic film.